粘果山羊草在小麦育种中的初步应用

为了将粘果山羊草的优异农艺性状转移到小麦背景中,本研究用(粘果山羊草/中国春)杂交F1代与普通小麦亲本复交并自交,共获得了42个F4代单株及其衍生的42个F5代株系,并从主要农艺性状、细胞学和高分子量谷蛋白亚基(HMW-GS)等方面对这一批材料进行了研究。研究发现:(1)F5代株系在形态上与普通小麦十分相似,部分株系的分蘖数和条锈病抗性有显著提高,部分株系的千粒重表现出超亲现象;(2)12个F4代植株的细胞学鉴定发现,7个单株的染色体数目与普通小麦不一致,其中1个单株还可能存在不稳定结构变异。这表明部分参试单株存在细胞学不稳定性现象,需要进一步自交纯合和鉴定;(3)聚丙烯酰胺凝胶电泳(SDS-PAGE)检测发现除1株变异株J7-4外,其余材料的HMW-GS组成均和普通小麦亲本一致。本研究为小麦育种提供了一批优异的育种材料。     

粘果山羊草在小麦改良中的初步应用

为了将粘果山羊草的优异农艺性状转移到小麦背景中,本研究用(粘果山羊草/中国春)杂交F1代与普通小麦亲本复交并自交,共获得了42个F4代单株及其衍生的42个F5代株系,并从主要农艺性状、细胞学和高分子量谷蛋白亚基(HMW-GS)等方面对这一批材料进行了研究。研究发现：1、上述F5代株系在形态上与普通小麦十分相似,部分株系的分蘖数和条锈病抗性有显著提高, 部分株系的千粒重表现出超亲现象,具有一定的育种利用价值;2、12个F4代植株的细胞学鉴定发现：7个单株的染色体数目与普通小麦不一致,其中1个单株还可能存在不稳定结构变异。这表明部分参试单株存在细胞学不稳定性现象,需要进一步自交纯合和鉴定;3、聚丙烯酰胺凝胶电泳(SDS-PAGE)检测发现除1株变异株J7-4外,其余材料的HMW-GS组成均和普通小麦亲本一致,变异株的变异机理和它对品质的影响还有待进一步深入研究。本研究为小麦育种提供了一批优异的育种材料。     

基于SSR分子标记技术的葡萄种间杂交F1代鉴定

SSR分子标记技术以其多态性高、重复性好和检测方便快捷等优点,被广泛应用于植物F1代真假杂种的鉴定。为了长期有效的开展葡萄杂交育种工作,本研究以葡萄杂交组合‘北冰红’伊‘贵人香’F1代159个单株及‘双红’伊‘贵人香’F1代121个单株为试验材料,利用SSR分子标记技术结合田间形态学性状对葡萄杂交后代的真伪进行鉴定。在12对SSR引物中筛选出同时在亲本间具有特异性位点的引物,对F1代进行扩增检测,结果表明:‘北冰红’伊‘贵人香’及‘双红’伊‘贵人香’2个杂交组合中分别有63和94个单株具有双亲特异性条带,结合田间形态学鉴定,认为其为真杂种,故2个杂交群体的杂种率分别为40.38%和77.69%,该结果可为今后开展葡萄遗传连锁图铺构建及数量性状的QTL定位提供依据。     

60Coγ诱变豌豆M2农艺性状与产量多重分析

为了更好的发掘利用豌豆辐射诱变变异后代资源,本研究分别以3种不同复叶类型豌豆为材料,通过对豌豆诱变M2代农艺性状进行相关分析、多元分析、通径分析、主成分分析,明确豌豆诱变后代产量的决定因子。结果表明不同复叶类型豌豆诱变M2的主要农艺性状的变异丰富,变异系数达0.171-0.8539;且结荚节数、单株荚数、单株粒数均与单株产量呈极显著正相关关系;通径分析结果证实,不同复叶类型豌豆诱变后代选择中,单株粒数及单株荚数对单株产量的影响均很大;主成分分析结果表明：单株荚数、单株粒数、结荚节数是高产型豌豆形态决定因子。豌豆诱变后代单株产量的主要决定因子依次为单株粒数、单株荚数、结荚节数,同时对百粒重给予足够重视。     

异源转座子存在的证据

本研究选用130对小麦SSR引物对从5个小麦-黑麦易位系中选取的56个单株进行PCR扩增,中国春,绵阳11和黑麦作为对照。结果发现122-2A和636-2A引物对在小麦和黑麦间产生多态性。122-2A引物在黑麦中产生2个黑麦特异条带,一个条带大小为190bp,另一个为330bp。在56个单株中有4个单株产生相应的黑麦特异带。这可能是黑麦中1个可移动因子(异源转座子)导入了小麦所致。并且结合许多病理学、进化生物学及其遗传学的证据对异源转座子在原核和真核生物中广泛存在的可能性进行了讨论。     

小麦未减数配子基因的连锁标记及染色体区段检测

六倍体普通小麦(Triticumaestivum L., AABBDD, 2n=42)由四倍体小麦(T. turgidum, AABB, 2n=28)与节节麦(Aegilops tauschii Cosson, DD, 2n=14)天然杂交,然后通过染色体自动加倍形成。加倍过程主要受四倍体小麦未减数配子基因控制,且不同四倍体小麦存在不同的遗传效应。本研究利用位于3B染色体上未减数配子基因QTug.sau-3B的连锁SSR标记Xgpw1146和高通量DArTseq分子标记,筛选出可能转入四倍体小麦未减数配子基因的人工合成小麦改良后代。在105份改良材料中检测出17份具有四倍体小麦的Xgpw1146等位位点,表明四倍体小麦的未减数配子基因可能转入了这17份材料。利用DArTseq高通量标记技术分析人工合成小麦SHW-L1的88份改良后代,发现含四倍体小麦Xgpw1146等位位点的材料均具有来自SHW-L1、且可能包含Xgpw1146的一个染色体区段,表明未减数配子基因临近区域以一个区段传递到改良后代。这些人工合成小麦改良材料在加倍单倍体育种中有重要的应用潜力。     

两个抗旱小麦资源主要农艺性状配合力和遗传力分析

本研究选用２个抗旱小麦和４个农艺亲本，按Ｇｒｉｆｉｎｇ双列杂交方法４配制１５个组合。对２个抗旱小麦Ｃ抗８和Ｃ抗９和１０个主要农艺性状的配合力和遗传进行了分析。结果表明：抗旱小麦Ｃ抗８在单株穗数、单株产量和收获指数等性状有高的一般配合力，且收获指数的特殊配合力方差最大。Ｃ抗９的每穗粒数、千粒重、单株产量和收获指数等几个性状的一般配合力效应值为负值，但其特殊配合力方差较高或中等。株高、千粒重、穗长、小     

八倍体小偃麦和硬粒小麦杂交后代的染色体组成分析

在小麦育种工作中,因长穗偃麦草、中间偃麦草和四倍体硬粒小麦等小麦近缘种属含有许多重要的功能基因,育种家经常应用远缘杂交创制小麦育种中间材料。本研究应用FISH、GISH、Mc-GISH技术检测了八倍体小偃麦和四倍体硬粒小麦杂交的后代材料,结果表明:山农20和四倍体硬粒小麦的杂交后代中,D组染色体显著优先于十倍体长穗偃麦草染色体传递到子代中;中3和中4与四倍体硬粒小麦的杂交后代中,D组和中间偃麦草染色体从1～14条随机传递到子代中;所有材料中仅从山农20和四倍体硬粒小麦的杂交后代中筛选出3份稳定的代换系,对其中的2576-1代换系进一步分析证明,是十倍体长穗偃麦草染色体代换了1D染色体并确定该材料抗条锈病,可以作为育种材料,也为异源新种质的创制奠定了基础。     

小麦抗白粉病基因Pm21分子标记辅助选择的应用

白粉病是威胁我国小麦生产最主要的病害之一。筛选和培育抗病品种是防治小麦白粉病的一条安全有效途径。Pm21是目前最有效的抗白粉病主效基因之一,加快其在小麦育种中的应用,对我国小麦生产具有重要的意义。本研究目的在于将普通小麦——簇毛麦易位系92R137中的抗白粉病基因Pm21导入农艺性状好、产量较高、较易感白粉病的农大系列小麦品种中。在92R137与农大系列小麦品种回交一代利用Pm21的SCAR1265标记进行检测,筛选出含有抗白粉病基因Pm21的单株,进行辅助育种应用的研究,经两次回交和两次标记辅助选择,从农大3291/92R137//农大32912组合后代中选出具有Pm21基因且产量性状好的品系13个,从农大3214/92R137//农大32142组合选出9个,从农大3213/92R137//农大32132组合选出35个,从农大3197/92R137//农大31972组合选出8个,从农大3383/92R137//农大33832组合选出15个,从农大3308/92R137//农大33082组合选出2个共计82个进入初步产量鉴定,根据初步产量结果和农艺性状表现选出9个优系参加2005年秋播产量试验。     

色素万寿菊产量构成因素的灰色关联度分析

运用灰色关联度分析方法,对10个色素万寿菊杂交组合的单株产量和7个主要性状(株高、株幅、茎粗、分枝数、花径、单株花数、单花重)进行灰色关联度分析.以研究各农艺性状对产量的影响.结果表明,单株花数、单花重、花径对单株产量的影响较大,结果与生产情况基本一致,说明在色素万寿菊育种后代选择上应将这几个性状作为主要指标.     

Development of dominant nuclear male-sterile lines with a blue seed marker in durum and common wheat

In order to develop genie male‐sterile lines with a blue seed marker, male‐sterile plants, controlled by a dominant nuclear gene Ms2, were used as female parents against a 4E disomic addition line ‘Xiaoyan Lanli’(2n= 44, AABBDD+4EII) as the male parent to produce monosomic addition lines with blue seed. Male‐sterile plants from the monosomic addition lines were pollinated with durum wheat for several generations and in 1989 a male‐sterile line with the blue grain gene and the male‐sterile gene Ms2 on the same additional chromosome was detected and named line 89‐2343. Using this line, the blue seed marker was successfully added to a short male‐sterile line containing Ms2 and Rht10. The segregation ratios of male sterility and seed colour as well as the chromosome figurations of different plants indicated that the blue grain genes, Ms2 and Rht10 were located on the same additional chromosome. Cytological analysis showed that the blue marker male‐sterile lines in durum wheat and common wheat were monosomic with an additional chromosome 4E. The inheritance ratio for blue seed male‐sterile plants and white seed male‐fertile plants was 19.7% and 80.3%, respectively, in common wheat. The potential for using blue marker sterile lines in population improvement and hybrid production is discussed.     

抗白粉病普通小麦-簇毛麦新种质的遗传学及生化鉴定

普通小麦-簇毛麦抗白粉病新种质94G22-1和94-G33-1都对白粉病免疫。其体细胞染色体数为42,花粉母细胞减数分裂中期Ⅰ染色体构型为21个二价体。用感病小麦品种对其测交,F_1代全部对白粉病表现免疫,F_1代花粉母细胞减数分裂中期Ⅰ染色体构型为20个二价体加2个单价体。测交F_2抗感比符合3:1的预期分离比例。体细胞快速荧光原位杂交表明94G22-1、94G33-1都带有两条簇毛麦染色体;染色体C-分带证明这两条簇毛麦染色体是一对6V;重双端体分析确认6V代换的普通小麦染色体是6D。在谷草转氨酶(GOT)同工酶电泳图潜的GOT-2活性区,94G22-1、94G33-1显示了3条酶带,其中1条来自簇毛麦,由6V上的基因编码,亚基组成分析亦确证94G22-1、94G33-1是6D(6V)异代换系。     

Effects of Zinc Deficiency and Drought on Grain Yield of Field-grown Wheat Cultivars in Central Anatolia

Drought stress and zinc (Zn) deficiency are serious abiotic stress factors limiting crop production in Turkey, especially in Central Anatolia. In this study, the effects of Zn deficiency and drought stress on grain yield of 20 wheat cultivars (16 bread wheat, Triticum aestivum; four durum wheat, Triticum durum cultivars) were investigated over 2 years under rainfed and irrigated conditions in Central Anatolia where drought and Zn deficiency cause substantial yield reductions. Plants were treated with (+Zn: 23 kg Zn ha⁻¹, as ZnSO₄·7H₂O) and without (−Zn) Zn under rainfed and irrigated conditions. Both Zn deficiency and rainfed treatments resulted in substantial decreases in grain yield. Significant differences were determined between both bread wheat and durum wheat cultivars in terms of drought stress tolerance. Considering drought sensitivity indices over 2 years, the bread wheat cultivars Yayla‐305, Gerek‐79, Dagdas‐94 and Bolal‐2973 were found to be more drought‐tolerant than the other cultivars under both −Zn and +Zn treatments. Especially the durum wheat cultivars Cakmak 79 and Selcuklu 97 showed much greater drought susceptibility under Zn deficiency, and irrigation alone was not sufficient to obtain satisfying grain yield without Zn application. The results indicate that sensitivity to Zn deficiency stress became more pronounced when plants were drought‐stressed. The effect of irrigation on grain yield was maximized when Zn was adequately supplied, leading to the suggestion that efficient water use in Central Anatolia seems to be highly dependent on the Zn nutritional status of plants.     

Chromosomal regions associated with the in vitro culture response of wheat (Triticum aestivum L.) microspores

Generation of doubled haploid plants is a powerful tool in breeding, as homozygous individuals will be obtained directly from hybrids. However, genotype variability in regeneration efficiency of most European wheat (Triticum aestivum L.) varieties has limited its use in wheat. This study intended to identify quantitative trait loci (QTLs) for green plantlet regeneration from wheat microspore cultures. A QTL analysis using DArT markers was conducted based on a bi‐parental F₃ population, derived from a cross between the varieties Svilena and Jensen, which displayed markedly different capacity for plantlet regeneration. Two QTLs on chromosome 1B and 7B explained 53% of the variation in green plantlet regeneration. Furthermore, a collection of 94 European wheat varieties was genotyped and phenotyped. The microspore response level was low among western and northern European wheat varieties, and the positive QTLs found in the bi‐parental population were rare in the variety collection. Identification of the two QTLs enables introduction of high regeneration efficiency into wheat germplasm. Moreover, our results proved that the efficient regeneration observed for one variety could be crossed into modern winter wheat.     

蓝标型小麦核雄性不育、保持系的选育研究

将蓝粒小麦中携带有蓝色胚乳基因和育性恢复基因的4E 染色体,附加到小麦核型雄性不育系上,选育出蓝标型小麦雄性不育、保持系。该体系浅蓝粒种子长出植株自交结实,粒色分离为深蓝、浅蓝和白粒。其中,白粒种子长出植株,全部为雄性不育,为普通小麦品种所恢复,杂种优势显著;浅蓝粒种子植株则自交结实,粒色仍分离为深蓝、浅蓝和白     

Effect of mannitol pretreatment to improve green plant regeneration on isolated microspore culture in Triticum turgidum ssp. durum cv. 'Jennah Khetifa'

The use of doubled haploids improves the efficiency of cultivar development in many crops and can be helpful in genetic and molecular studies. The major problem with this approach is the low efficiency of green plant regeneration. We describe here an efficient method for inducing embryos and regenerating green plants directly from isolated microspores of durum wheat cv. ‘Jennah Khetifa’. Tillers from donor plants were pretreated in 0.3 m mannitol and were stored at 4°C at various times: 3, 5, 6, 7, 8, 10 and 12 days. Our results showed clearly that the novel pretreatment combined mannitol 0.3 m and cold for 7 days had a strong effect on the number of embryos produced and regenerated green plants. Under this condition 13 475 mature embryos were produced from 2 693 500 microspores. Moreover, 85 green plants were obtained. High green plants regeneration frequency was recorded. As an average 11.55 green plants were produced per 100 000 microspores (about the equivalent of six plants per spike). Therefore, this study showed clearly that our results are the best ones published until now in durum wheat.     

根癌农杆菌介导获得稗草Ecppc转基因小麦的研究

采用携带pUbi-Ecppc质粒的3个根癌农杆菌菌株（LBA4404、EHA105和C58c1）,对经过预培养10~12 d的春小麦品种扬麦158、Bobwhite和扬麦12的幼胚愈伤组织进行了遗传转化。对筛选中的抗生素浓度、菌液浓度、共培养温度和时间、受体基因型、菌株-质粒组合等影响转化的重要因素进行了研究。首次将单子叶野生C₄植物稗草的磷酸烯醇式丙酮酸羧化酶基因（Ecppc）导入小麦受体基因型,并得到具有潮霉素（Hyg）抗性的转化植株。从816块共培养愈伤组织中转化得到34株抗性植株,其中14株PCR检测为阳性。扬麦158的转化效率达3.03%。Southern和RT-PCR分析表明外源基因已整合到小麦基因组并得到正确的转录。生理学检测显示,转基因小麦植株的光合速率和PEPC活性都有所提高。说明Ubiqintin基因启动子控制的稗草PEPC cDNA基因在小麦中可以正确表达和起到一定的生理作用。这些工作为进一步探讨PEPC对小麦光合作用及其他生理过程的影响奠定了基础。     

抗纹枯病、根腐病的转SN1基因小麦的获得与鉴定

SN1是源于马铃薯的一种抗菌肽, 可以抑制多种植物病原菌的生长。小麦纹枯病(主要病原菌为禾谷丝核菌Rhizoctonia cerealis)和根腐病(主要病原菌为平脐蠕孢菌Bipolaris sorokiniana)是小麦的主要土传真菌病害。本研究利用基因工程技术构建了SN1基因的单子叶植物表达载体pA25-SN1, 它受玉米泛素(ubiquitin)启动子的控制;采用基因枪法将pA25-SN1转化小麦推广品种扬麦18幼胚愈伤组织4 000块, 获得203株再生植株, 通过PCR检测出阳性植株55株, 转化率为1.38%。对转SN1基因小麦T₀~T₂代植株, 进行外源基因的PCR、Southern blot、RT-PCR、荧光定量RT-PCR(Q-RT-PCR)分析和小麦纹枯病菌与根腐病菌接种及其抗病性鉴定。结果表明, 转入的SN1基因已经整合到转基因小麦的基因组中, 能够在转基因小麦中遗传、转录与表达。SN1基因的表达提高了转基因植株对小麦纹枯病和根腐病的抗性, 其抗病性可以遗传。     

转Rs-AFP2基因小麦的分子分析及其纹枯病抗性

Rs-AFP2属于r-硫堇类抗菌肽,主要通过形成离子通道直接破坏细胞来杀灭病原菌。本研究通过基因枪介导法结合对目标基因的分子检测,证明已将外源Rs-AFP2基因转入小麦推广品种扬麦12中。通过逐株抗纹枯病接种鉴定、PCR、PCR-Southern blot、Southern blot和 RT-PCR/荧光定量RT-PCR(Q-RT-PCR)分析,对转Rs-AFP2基因小麦T₁至T₄代植株跟踪检测。结果表明,Rs-AFP2在转基因小麦中能够稳定遗传,以单拷贝整合到小麦基因组中,遗传方式符合孟德尔遗传规律,并能在转录水平上表达。对转Rs-AFP2基因小麦的抗病性、主要农艺性状以及Rs-AFP2表达活性分析结果表明,与受体扬麦12相比,Rs-AFP2表达活性高的转基因小麦植株对纹枯病抗性有明显提高,其抗病性可以遗传,而主要农艺性状没有明显差异,证明可以利用Rs-AFP2基因和基因工程途径创制抗纹枯病小麦新种质。